CN107460148A - A kind of preparation method and application of bacillus subtilis bacterium powder - Google Patents
A kind of preparation method and application of bacillus subtilis bacterium powder Download PDFInfo
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- CN107460148A CN107460148A CN201710779910.8A CN201710779910A CN107460148A CN 107460148 A CN107460148 A CN 107460148A CN 201710779910 A CN201710779910 A CN 201710779910A CN 107460148 A CN107460148 A CN 107460148A
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- bacillus subtilis
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- bacterium powder
- subtilis bacterium
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- 240000008371 Bacillus subtilis Species 0.000 title claims abstract description 75
- 239000000843 powder Substances 0.000 title claims abstract description 59
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 241000894006 Bacteria Species 0.000 claims abstract description 78
- 239000007788 liquid Substances 0.000 claims abstract description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 35
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 28
- 229940075615 Bacillus subtilis Drugs 0.000 claims abstract description 27
- 238000000855 fermentation Methods 0.000 claims abstract description 15
- 230000004151 fermentation Effects 0.000 claims abstract description 15
- 238000005119 centrifugation Methods 0.000 claims abstract description 14
- 239000003223 protective agent Substances 0.000 claims abstract description 11
- 229940073490 Sodium Glutamate Drugs 0.000 claims abstract description 10
- HDTRYLNUVZCQOY-LIZSDCNHSA-N Trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 10
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims abstract description 10
- LPUQAYUQRXPFSQ-UHFFFAOYSA-M monosodium glutamate Chemical compound [Na+].[O-]C(=O)C(N)CCC(O)=O LPUQAYUQRXPFSQ-UHFFFAOYSA-M 0.000 claims abstract description 10
- 235000013923 monosodium glutamate Nutrition 0.000 claims abstract description 10
- GUBGYTABKSRVRQ-UUNJERMWSA-N Lactose Natural products O([C@@H]1[C@H](O)[C@H](O)[C@H](O)O[C@@H]1CO)[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1 GUBGYTABKSRVRQ-UUNJERMWSA-N 0.000 claims abstract description 9
- GUBGYTABKSRVRQ-XLOQQCSPSA-N lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims abstract description 9
- 239000008101 lactose Substances 0.000 claims abstract description 9
- 239000002504 physiological saline solution Substances 0.000 claims abstract description 9
- 239000010841 municipal wastewater Substances 0.000 claims abstract description 6
- 238000004108 freeze drying Methods 0.000 claims abstract description 5
- 239000010842 industrial wastewater Substances 0.000 claims abstract description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- 239000012530 fluid Substances 0.000 claims description 4
- 239000012266 salt solution Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 4
- 230000014759 maintenance of location Effects 0.000 abstract description 2
- 238000003756 stirring Methods 0.000 description 12
- 239000010865 sewage Substances 0.000 description 7
- 239000002351 wastewater Substances 0.000 description 7
- 241000193830 Bacillus <bacterium> Species 0.000 description 6
- 238000011084 recovery Methods 0.000 description 6
- CVTZKFWZDBJAHE-UHFFFAOYSA-N [N].N Chemical compound [N].N CVTZKFWZDBJAHE-UHFFFAOYSA-N 0.000 description 5
- 230000037250 Clearance Effects 0.000 description 4
- 230000035512 clearance Effects 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 239000003344 environmental pollutant Substances 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- NPDODHDPVPPRDJ-UHFFFAOYSA-N permanganate Chemical compound [O-][Mn](=O)(=O)=O NPDODHDPVPPRDJ-UHFFFAOYSA-N 0.000 description 4
- 231100000719 pollutant Toxicity 0.000 description 4
- 238000004065 wastewater treatment Methods 0.000 description 4
- 241000276438 Gadus morhua Species 0.000 description 3
- 241000726221 Gemma Species 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 235000019516 cod Nutrition 0.000 description 3
- 239000002270 dispersing agent Substances 0.000 description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 239000011574 phosphorus Substances 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000004563 wettable powder Substances 0.000 description 3
- 210000000481 Breast Anatomy 0.000 description 2
- JBKVHLHDHHXQEQ-UHFFFAOYSA-N Caprolactam Chemical compound O=C1CCCCCN1 JBKVHLHDHHXQEQ-UHFFFAOYSA-N 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 2
- 210000002784 Stomach Anatomy 0.000 description 2
- 230000001580 bacterial Effects 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000004059 degradation Effects 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- MYMOFIZGZYHOMD-UHFFFAOYSA-N oxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000010802 sludge Substances 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 1
- 210000000936 Intestines Anatomy 0.000 description 1
- XUWPJKDMEZSVTP-UHFFFAOYSA-N Kalamycin Chemical compound O=C1C2=C(O)C=CC=C2C(=O)C2=C1C(C)OC1C2OC(=O)C1 XUWPJKDMEZSVTP-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000108664 Nitrobacteria Species 0.000 description 1
- ZJBYBXHCMWGGRR-UHFFFAOYSA-M Permanganic acid Chemical class O[Mn](=O)(=O)=O ZJBYBXHCMWGGRR-UHFFFAOYSA-M 0.000 description 1
- -1 Propylhomoserin sodium Chemical compound 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 229940088594 Vitamin Drugs 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating Effects 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000005276 aerator Methods 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 102000004139 alpha-Amylases Human genes 0.000 description 1
- 108090000637 alpha-Amylases Proteins 0.000 description 1
- 229940024171 alpha-amylase Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000003416 augmentation Effects 0.000 description 1
- 244000052616 bacterial pathogens Species 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 239000010866 blackwater Substances 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000005446 dissolved organic matter Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 150000004676 glycans Polymers 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 239000001963 growth media Substances 0.000 description 1
- 238000005338 heat storage Methods 0.000 description 1
- 239000003906 humectant Substances 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000000813 microbial Effects 0.000 description 1
- 230000002906 microbiologic Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 244000005706 microflora Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 238000005453 pelletization Methods 0.000 description 1
- 230000000243 photosynthetic Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 235000019422 polyvinyl alcohol Nutrition 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 210000004215 spores Anatomy 0.000 description 1
- 230000028070 sporulation Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000002352 surface water Substances 0.000 description 1
- 230000004083 survival Effects 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 229930003231 vitamins Natural products 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
- C02F2101/206—Manganese or manganese compounds
Abstract
The invention discloses a kind of preparation method and application of bacillus subtilis bacterium powder, the preparation method includes：Bacillus subtilis fermentation liquor is taken, bacteria residue is collected by centrifugation；Physiological saline and protective agent are added into bacteria residue, obtains mixed bacteria liquid；On the basis of mixed bacteria liquid, the protective agent includes following components：5~10wt% of trehalose, 5~10wt% of lactose, 0.5~3wt% of sodium glutamate；After pre-freeze, freeze-drying and crushing, bacillus subtilis bacterium powder is made successively in mixed bacteria liquid.The bacillus subtilis bacterium powder high income that preparation method of the present invention obtains, living bacteria count is high, retention cycle length, and can more effectively improve the treatment effect of municipal wastewater, industrial wastewater, black and odorous water and natural river water body.
Description
Technical field
The present invention relates to technical field of environmental pollution treatment, more particularly to a kind of preparation method of bacillus subtilis bacterium powder
And application.
Background technology
Bacillus subtillis is one kind in bacillus, is to have mesophilic, aerobic, sporiferous bacillus, its
Physiological characteristic is various, widely distributed, is especially easily separately cultured in soil sample.Bacillus stability is good, resistance, tool
There is the features such as high temperature resistant, acid and alkali-resistance, anti-extrusion, in industrial production pelletization and pass through cultivated animals acidic stomach environment
When, it can keep the stability of height.After the sporulation of bacillus subtilis, various unfavorable conditions are resistant to.
Currently, bacillus is fully developed, such as：Improve animal intestines and stomach work(as feed addictive
Energy；Improve water quality in terms of aquaculture, improve aquatic livestock survival rate；In terms of plantation, crop yield etc. is improved.
But in terms of sewage disposal, it is more than the utilization relatively of bacillus several inferior many.In recent years, gemma
Research on utilization of the bacillus in terms of sewage is in the ascendant, and bacillus subtilis being capable of intensive decomposition carbon system, nitrogen system, phosphorus system, sulphur system
Deng pollutant, dissolved organic matter in the polysaccharide, protein and water of complexity can be decomposed, turns into dominant microflora in water.
At present, bacillus subtilis has turned into the focus of water purification research.Bacillus subtilis can breed rapidly in water body,
Substantial amounts of ectoenzyme is produced, it is a large amount of to consume COD organic matters, reduce water pollution.Bacillus subtilis can be by larger molecular organicses
Be decomposed into the materials such as organic acid, amino acid and carbon dioxide and the water of small molecule, can be water body in nitrobacteria, photosynthetic bacteria,
The beneficial microbes such as lactic acid bacteria, algae and other planktonic organisms provide nutrition, the energy for having certain degradation of ammonia nitrogen, nitrite
Power.
Bacillus subtilis bacterium powder is a kind of product form, is the concentrate after fermentation centrifuges, with the addition of certain
Protective agent and dispersant so that bacillus subtilis more preferable storage and transport under solid forms, reduce production cost, carry
The high value of bacillus subtilis.
Currently, it is too numerous to enumerate on the technical study of bacillus subtilis bacterium powder and patent application.Such as：
Zheng Yong profits wait (Zheng Yongli, thunder beautiful eyes bacillus subtilis powders agent grinding as chicken feed addictive within (1) 1998 year
Study carefully Zhongshan University journal natural science edition, 02 phase in 1998) using with kalamycin resistance and the life that alpha-amylase can be secreted
Thing engineering bacteria Bacillus subtilis Bs964 adds pulvis as microbiological feed.
(Wang Jian, Wang Nan, Gao Guanpeng, Yin Danhan, hundred million gemma of Wang Wei .200/g bacillus subtilises such as (2) 2010 years Wang Jian
The development agricultural chemicals of wettable powder, 49 phases in 2010) using bacillus subtilis B99-2 as research object, by filler, profit
The protectant screening of humectant, dispersant, stabilizer, UV, it is determined that the optimum formula of bacillus subtilis wettable powder.This is most
Good formula is filler diatomite 10%, dispersant PVA 7.2%, wetting agent D425 4.8%, stabilizer CMC-Na 2.0%, is protected
Protect agent FWA 0.1%.Measurement result shows：The spore content of bacillus subtilis wettable powder is up to 2.0 × 1010Cfu/g, hang
Floating rate 79%, wetting time 48s, heat storage resolution ratio 23.6%, product indices are up to state standards.
(3) Chinese patent literature of Application No. 201310219751.8 discloses a kind of system of bacillus subtilis bacterium powder
Preparation Method, the invention pass through the exploration to culture medium prescription and fermentation condition, the strong bacterium powder for solving conventional method preparation
The problem of viable count content is few, gemma conversion ratio is low, but the preparation technology of bacterium powder is not optimized, do not obtain more
Efficient bacterium powder prepares data.
Therefore, it is necessary to the preparation method of new bacillus subtilis bacterium powder is deeply probed into again.
The content of the invention
The invention provides a kind of preparation method and application of bacillus subtilis bacterium powder, is obtained using the preparation method
Bacillus subtilis bacterium powder high income, living bacteria count is high, retention cycle length, and can more effectively improve municipal wastewater, work
The treatment effect of industry waste water, black and odorous water and natural river water body.
A kind of preparation method of bacillus subtilis bacterium powder, including：
(1) bacillus subtilis fermentation liquor is taken, bacteria residue is collected by centrifugation；
(2) physiological saline and protective agent are added into bacteria residue, obtains mixed bacteria liquid；
On the basis of mixed bacteria liquid, the dosage of the bacteria residue is 67.0~84.5wt%, and the protective agent includes following
Component：5~10wt% of trehalose, 5~10wt% of lactose, 0.5~3wt% of sodium glutamate；
(3) bacillus subtilis bacterium powder is made successively after pre-freeze, freeze-drying and crushing in mixed bacteria liquid.
Specifically, the deposit number of the bacillus subtilis is CCTCC M 2015688.
Preferably, in step (1), after adding diatomite into the zymotic fluid, then centrifuged；Using zymotic fluid as base
Standard, the diatomaceous dosage are 1~2.5wt%.Diatomaceous addition can more efficiently prevent from damage of the centrifugation to bacterial strain
Wound, improve viable count.
Preferably, in step (2), on the basis of mixed bacteria liquid, the dosage of the physiological saline is 5~10wt%；
The addition of physiological saline can be such that bacteria residue and protective agent fully mixes.
Further preferably, on the basis of mixed bacteria liquid, the protective agent, including following components：Trehalose 10wt%, lactose
10wt%, sodium glutamate 3wt%.
Preferably, in step (2), the temperature of the pre-freeze is -20~-40 DEG C, and the time is 48~96h.
Preferably, in step (2), the temperature of the freeze-drying is -45~-55 DEG C, and the time is 48~72h.
The invention provides bacillus subtilis bacterium powder prepared by a kind of described preparation method, the outward appearance of the bacterium powder is light
Yellow powder, the solubility in water are 50~100g/L, and viable count CFU is 7.22 × 1011~1.0 × 1012/g。
Present invention also offers described bacillus subtilis bacterium powder in processing municipal wastewater, industrial wastewater, black and odorous water
Or the application in natural river water body.
Especially in the chemical wastewater treatment of high salinity and high concentration, bacillus subtilis bacterium powder of the present invention is shown
High-salt tolerance, it can effectively reduce the CODcr in waste water.
Further, described application, on the basis of water, the dosage of the bacillus subtilis bacterium powder is 0.1ppm
~10ppm.
Compared with prior art, the invention has the advantages that：
The present invention carries out the preparation of bacillus subtilis bacterium powder using specific components and the protective agent of dosage, and coordinates suitably
Preparation technology, obtained high income and be up to 7.22 × 10 up to 50~60%, viable count CFU11~1.0 × 1012/ g, preserve week
Phase grows the bacillus subtilis bacterium powder of 12~18 months, and by the bacillus subtilis bacterium powder be used for municipal wastewater, industrial wastewater,
In black and odorous water or natural river water body so that water body major pollutants ammonia nitrogen, total nitrogen, permanganate index etc. decline 40~
90%, wastewater treatment efficiency is notable.
Embodiment
The bacillus subtilis strain being related in the following example is the preservation of China typical culture collection center preservation
Numbering is CCTCC M 2015688 bacterial strain；The present invention is not related to the preservation of strain.
Embodiment 1
A kind of preparation method of bacillus subtilis bacterium powder, is comprised the following steps that：
(1) bacillus subtilis fermentation liquor for taking deposit number to be CCTCC M 2015688, adds 1.5wt% thereto
Diatomite (on the basis of bacillus subtilis fermentation liquor), after stirring, 300s is centrifuged at 10000r/min, 25 DEG C,
After centrifugation terminates, bacteria residue is collected；
(2) bacteria residue (on the basis of mixed bacteria liquid, mass fraction 81wt%) for being collected by centrifugation to obtain is taken, is added thereto
6wt% physiological saline (on the basis of mixed bacteria liquid), after stirring, add 6wt% trehalose, 6wt% lactose,
1wt% sodium glutamate (on the basis of mixed bacteria liquid), stirs to bacteria residue after being evenly distributed, obtains mixed bacteria liquid；
(3) mixed bacteria liquid is placed in -20 DEG C of pre-freeze 96h, then 48h is freeze-dried at -55 DEG C, crushed 50 mesh sieves, obtain
To bacillus subtilis bacterium powder.
Total recovery is 53.21%, and the viable count CFU of bacillus subtilis bacterium powder is 7.22 × 1011/ g, under the conditions of 10 DEG C
Holding time is 18 months.
Embodiment 2
A kind of preparation method of bacillus subtilis bacterium powder, is comprised the following steps that：
(1) bacillus subtilis fermentation liquor for taking deposit number to be CCTCC M 2015688, adds 2.5wt% thereto
Diatomite (on the basis of bacillus subtilis fermentation liquor), after stirring, 300s is centrifuged at 10000r/min, 25 DEG C,
After centrifugation terminates, bacteria residue is collected；
(2) bacteria residue (on the basis of mixed bacteria liquid, mass fraction 68wt%) for being collected by centrifugation to obtain is taken, is added thereto
10wt% physiological saline (on the basis of mixed bacteria liquid), after stirring, add 10wt% trehalose, 10wt% breast
Sugar, 3wt% sodium glutamate (on the basis of mixed bacteria liquid), stir to bacteria residue after being evenly distributed, obtain mixed bacteria liquid；
(3) mixed bacteria liquid is placed in -40 DEG C of pre-freeze 48h, then 72h is freeze-dried at -55 DEG C, 50 mesh sieves are crossed after crushing,
Obtain bacillus subtilis bacterium powder.
Total recovery is 58.11%, and the viable count CFU of bacillus subtilis bacterium powder is 1.0 × 1012/ g, at ambient temperature
Holding time is 12 months.
Embodiment 3
A kind of preparation method of bacillus subtilis bacterium powder, is comprised the following steps that：
(1) bacillus subtilis fermentation liquor for taking deposit number to be CCTCC M 2015688, adds 2.5wt% thereto
Diatomite (on the basis of bacillus subtilis fermentation liquor), after stirring, 300s is centrifuged at 10000r/min, 25 DEG C,
After centrifugation terminates, bacteria residue is collected；
(2) bacteria residue (on the basis of mixed bacteria liquid, mass fraction 68wt%) for being collected by centrifugation to obtain is taken, is added thereto
10wt% physiological saline (on the basis of mixed bacteria liquid), after stirring, add 10wt% trehalose, 10wt% breast
Sugar, 2wt% sodium glutamate (on the basis of mixed bacteria liquid), stir to bacteria residue after being evenly distributed, obtain mixed bacteria liquid；
(3) mixed bacteria liquid is placed in -40 DEG C of pre-freeze 48h, then 72h is freeze-dried at -55 DEG C, 50 mesh sieves are crossed after crushing,
Obtain bacillus subtilis bacterium powder.
Total recovery is 53.05%, and the viable count CFU of bacillus subtilis bacterium powder is 9.13 × 1011/ g, at ambient temperature
Holding time is 12 months.
Comparative example 1
(1) bacillus subtilis fermentation liquor for taking deposit number to be CCTCC M 2015688, adds 2.5wt% thereto
Diatomite (on the basis of bacillus subtilis fermentation liquor), after stirring, 300s is centrifuged at 10000r/min, 25 DEG C,
After centrifugation terminates, bacteria residue is collected；
(2) bacteria residue for being collected by centrifugation to obtain is taken, adds 10wt% physiological saline (on the basis of mixed bacteria liquid) thereto,
After stirring, trehalose, lactose, sodium glutamate (on the basis of mixed bacteria liquid, specific proportioning such as following table) are added, stirring is extremely
After bacteria residue is evenly distributed, mixed bacteria liquid is obtained；
(3) mixed bacteria liquid is placed in -40 DEG C of pre-freeze 48h, then 72h is freeze-dried at -55 DEG C, 50 mesh sieves are crossed after crushing,
Obtain that bacillus subtilis bacterium powder is made.
Specific bacterium powder viable count and total recovery such as following table：
| Scheme 1 | Scheme 2 | Scheme 3 | Scheme 4 | Scheme 5 | |
| Trehalose (wt%) | - | 10% | 10% | 15% | 3% |
| Lactose (wt%) | 10% | - | 10% | 15% | 3% |
| Sodium glutamate (wt%) | 3% | 3% | - | 5% | 0.3% |
| Bacterium powder viable count (CFU/g) | 1.7×1010 | 3.2×1010 | 7×1010 | 5.8×1010 | 1.0×1010 |
| Total recovery (%) | 5.51 | 11.00 | 39.78 | 44.56 | 2.39 |
Comparative example 2
(1) bacillus subtilis fermentation liquor for taking deposit number to be CCTCC M 2015688, adds 2.5wt% thereto
Diatomite (on the basis of bacillus subtilis fermentation liquor), after stirring, 300s is centrifuged at 10000r/min, 25 DEG C,
After centrifugation terminates, bacteria residue is collected；
(2) bacteria residue for being collected by centrifugation to obtain is taken, 75wt% distilled water (on the basis of mixed bacteria liquid) is added thereto, stirs
After mixing uniformly, 0.75wt% skimmed milk, 0.6wt% lactose, 0.15wt% vitamin Cs 1,0.075wt% paddy are added
Propylhomoserin sodium (on the basis of mixed bacteria liquid), stirs to bacteria residue after being evenly distributed, obtains mixed bacteria liquid；
(3) mixed bacteria liquid is freeze-dried, obtains lyophilized bacterium powder, viable count CFU is 3.0 × 10 in bacterium powder11/ g, total recovery
For 24.72%, the holding time is 12 months at room temperature.
Application examples 1
Bacillus subtilis bacterium powder prepared by above-described embodiment 2 is applied in the processing of municipal wastewater, and particular content is such as
Under：
The application example processing sewage source in Zhejiang municipal sewage plant, main sewage type based on dyeing waste water,
Its Biochemical Treatment is poor, and influent COD cr is up to 320mg/L, more than water outlet CODcr 60mg/L after its former PROCESS FOR TREATMENT, difficult
With qualified discharge.
Using its second stage of the project of function bacterium bacterium powder intensive treatment technical finesse, i.e., add 1ppm in the second stage of the project aeration tank
Bacillus subtilis bacterium powder, using strain energy efficient degradation COD characteristic, after 1 month, water outlet CODcr stably reaches continuous service
To below 50mg/L, reach《Urban wastewater treatment firm pollutant emission standard》GB18918-2002 one-level A standards, CODcr are gone
Except rate by 81.3% is promoted to 84.3%.
By contrast, its first stage of the project still fails to reach stably reaching standard row after handling by the way of bacterium solution directly adds
The effect put；The third stage of the project takes the bacterium powder prepared described in comparative example 2 to be handled, and its water quality is unable to qualified discharge.
Application examples 2
Bacillus subtilis bacterium powder prepared by above-described embodiment 2 is applied in river course reparation, and particular content is as follows：
Zhejiang garden river course, belong to industrial park typical case and receive dirty river course, quality of river water leading indicator ammonia nitrogen 3.2~
5.7mg/L, dissolved oxygen ＜ 2mg/L, 17~19mg/L of permanganate index, 0.58~0.61mg/L of total phosphorus, are chronically at earth's surface
The bad V classes water of water, eutrophication is more serious, and summer, the river course part waters river was smelly, and blue algae bloom is serious.
By to the river course suitable control exchange capacity and aerator being simply provided, river dissolving bacterium powder is taken directly to add
Mode, add 0.5ppm bacillus subtilises bacterium powder and carry out microbial augmentation improvement, by the debugging of one month, handle section
1.2~1.5mg/L of quality of river water leading indicator ammonia nitrogen, dissolved oxygen ＞ 7mg/L, 5~9mg/L of permanganate index, total phosphorus 0.28
~0.38mg/L, III~IV class water is improved as by the bad V classes water of surface water, and handled before this section of river course by other enterprises,
Its water quality is not significantly improved.
Application examples 3
Bacillus subtilis bacterium powder prepared by above-described embodiment 2 is applied in chemical wastewater treatment, and particular content is as follows：
For Quzhou chemical enterprise to produce based on caprolactam, the daily waste water that produces reaches 1000m3, comprehensive wastewater CODcr reaches
5000mg/L, electrical conductivity 20000us/cm, waste water B/C belong to the high salinity used water difficult to degradate of typical high concentration than 0.14.
There was only 50% or so using conventional activated sludge process processing CODcr clearances, and sludge bulking, microorganism often occur
Phenomena such as inactivation.
Using function bacterium intensive treatment technology, i.e., the bacillus subtilis bacterium of 2ppm concentration is added in biochemical reaction tank
Powder, then it is vexed expose culture 5 days after intake debugging, water outlet CODcr stably reaches below 500mg/L after debugging in 20 days, on clearance
90% is raised to, significantly improves sewage treating efficiency.
Illustrate that bacillus subtilis bacterium powder prepared by embodiment 2 has application excellent in high salinity and high-concentration waste water is handled
Gesture.The bacterium powder prepared described in comparative example 2 is taken equally to be handled, its effluent quality CODcr can not stably reach 500mg/L.
Application examples 4
Bacillus subtilis bacterium powder prepared by above-described embodiment 2 is applied in the reparation of malodorous black water body, and particular content is such as
Under：
, beyond the ability of the river self-purification, led because sanitary sewage is discharged into river course by neighbouring resident for a long time in Huzhou cities and towns inland river
Cause river water body to black smelly, turn into bad V classes water, permanganate index is up to 30mg/L or so, ammonia nitrogen 8mg/L or so.Pass through
1.5ppm bacillus subtilis bacterium powders are added, after processing in 10 days, each pollutant has obvious removal effect, wherein permanganic acid
Salt index is handled to≤12mg/L, clearance more than 60%, NH_3-N treating to≤1.8mg/L, clearance more than 77.5%.
Claims (10)
- A kind of 1. preparation method of bacillus subtilis bacterium powder, it is characterised in that including：(1) bacillus subtilis fermentation liquor is taken, bacteria residue is collected by centrifugation；(2) physiological saline and protective agent are added into bacteria residue, obtains mixed bacteria liquid；On the basis of mixed bacteria liquid, the dosage of the bacteria residue is 67.0~84.5wt%, and the protective agent includes following components： 5~10wt% of trehalose, 5~10wt% of lactose, 0.5~3wt% of sodium glutamate；(3) bacillus subtilis bacterium powder is made successively after pre-freeze, freeze-drying and crushing in mixed bacteria liquid.
- 2. preparation method as claimed in claim 1, it is characterised in that the deposit number of the bacillus subtilis is CCTCC M 2015688。
- 3. preparation method as claimed in claim 1, it is characterised in that in step (1), diatomite is added into the zymotic fluid Afterwards, then centrifuged；On the basis of zymotic fluid, the diatomaceous dosage is 1~2.5wt%.
- 4. preparation method as claimed in claim 1, it is characterised in that in step (2), on the basis of mixed bacteria liquid, the life The dosage for managing salt solution is 5~10wt%.
- 5. preparation method as claimed in claim 1, it is characterised in that on the basis of mixed bacteria liquid, the protective agent, including with Lower component：Trehalose 10wt%, lactose 10wt%, sodium glutamate 3wt%.
- 6. preparation method as claimed in claim 1, it is characterised in that in step (2), the temperature of the pre-freeze is -20~-40 DEG C, the time is 48~96h.
- 7. preparation method as claimed in claim 1, it is characterised in that in step (2), the temperature of the freeze-drying is -45 ~-55 DEG C, the time is 48~72h.
- A kind of 8. bacillus subtilis bacterium powder prepared by preparation method as claimed in claim 1.
- 9. bacillus subtilis bacterium powder as claimed in claim 8 is in processing municipal wastewater, industrial wastewater, black and odorous water or nature Application in river water body.
- 10. application as claimed in claim 9, it is characterised in that on the basis of water, the throwing of the bacillus subtilis bacterium powder Dosage is 0.1ppm~10ppm.
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| CN110468063A (en) * | 2019-04-17 | 2019-11-19 | 浙江亲水园生物科技有限公司 | A kind of microbial solid preparation, preparation method and application |
| CN111748497A (en) * | 2020-07-07 | 2020-10-09 | 山东农业大学 | Bacillus megaterium and application thereof in rapid degradation of nitrite |
| CN111748497B (en) * | 2020-07-07 | 2022-02-08 | 山东农业大学 | Bacillus megaterium and application thereof in rapid degradation of nitrite |
| CN112011486A (en) * | 2020-09-09 | 2020-12-01 | 天津市农业科学院 | Synchronous nitrification and denitrification bacillus subtilis K8 and application thereof |
| CN112011486B (en) * | 2020-09-09 | 2022-03-01 | 天津市农业科学院 | Synchronous nitrification and denitrification bacillus subtilis K8 and application thereof |
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